Del Prado M, Hernández-Montes H, Villalpando S
Unidad de Investigación en Nutrición, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, IMSS, México, D.F.
Arch Med Res. 1994 Autumn;25(3):331-5.
A fluorometric assay for determining lipoprotein lipase (LPL) activity is described. Dibutyrilfluorescine (DBF) was used as substrate for the enzyme and the fluoresceine liberated by enzymatic hydrolysis of the substrate was measured. Extracts of acetone powder from adipose tissue as an enzyme source showed characteristics of lipoprotein lipase activity, i.e., inhibition by NaCl and optimum activity in alkaline pH. There was close agreement in LPL activity when the same sample was measured simultaneously using either dibutyrilfluorescine or tri[9,10(3)H]oleylglycerol as substrate. The extent of inhibition of lipoprotein lipase by NaCl was similar with both methods. The fluorometric method detected changes in LPL activity in heart and adipose tissue related to the nutritional status of the animal with the same specificity and sensitivity than did the radioactive method. The fluorometric method is as sensitive, less expensive and less time consuming than the radioactive method.
本文描述了一种用于测定脂蛋白脂肪酶（LPL）活性的荧光测定法。使用二丁酰荧光素（DBF）作为该酶的底物，并测定底物经酶水解后释放的荧光素。来自脂肪组织的丙酮粉提取物作为酶源，显示出脂蛋白脂肪酶活性的特征，即受NaCl抑制且在碱性pH下具有最佳活性。当使用二丁酰荧光素或三[9,10(3)H]油酰甘油作为底物同时测定同一样品时，LPL活性结果高度一致。两种方法中NaCl对脂蛋白脂肪酶的抑制程度相似。荧光测定法检测到心脏和脂肪组织中LPL活性的变化与动物的营养状况相关，其特异性和敏感性与放射性方法相同。荧光测定法与放射性方法一样灵敏，但成本更低且耗时更少。
